Martin Lundqvist

Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts

Nanoparticles in a biological fluid (plasma, or otherwise) associate with a range of biopolymers, especially proteins, organized into the “protein corona” that is associated with the nanoparticle and continuously exchanging with the proteins in the environment. Methodologies to determine the corona and to understand its dependence on nanomaterial properties are likely to become important in bionanoscience. Here, we study the long-lived (“hard”) protein corona formed from human plasma for a range of nanoparticles that differ in surface properties and size. Six different polystyrene nanoparticles were studied: three different surface chemistries (plain PS, carboxyl-modified, and amine-modified) and two sizes of each (50 and 100 nm), enabling us to perform systematic studies of the effect of surface properties and size on the detailed protein coronas. Proteins in the corona that are conserved and unique across the nanoparticle types were identified and classified according to the protein functional properties. Remarkably, both size and surface properties were found to play a very significant role in determining the nanoparticle coronas on the different particles of identical materials. We comment on the future need for scientific understanding, characterization, and possibly some additional emphasis on standards for the surfaces of nanoparticles.

The Evolution of the Protein Corona around Nanoparticles: A Test Study

The importance of the protein corona formed around nanoparticles upon entering a biological fluid has recently been highlighted. This corona is, when sufficiently long-lived, thought to govern the particles biological fate. However, even this long-lived hard corona evolves and re-equilibrates as particles pass from one biological fluid to another, and may be an important feature for long-term fate. Here we show the evolution of the protein corona as a result of transfer of nanoparticles from one biological fluid (plasma) into another (cytosolic fluid), a simple illustrative model for the uptake of nanoparticles into cells. While no direct comparison can be made to what would happen in, for example, the uptake pathway, the results confirm that significant evolution of the corona occurs in the second biological solution, but that the final corona contains a fingerprint of its history. This could be evolved to map the transport pathways utilized by nanoparticles, and eventually to predict nanoparticle fate and behavior.

Modeling the Time Evolution of the Nanoparticle-Protein Corona in a Body Fluid

Background Nanoparticles in contact with biological fluids interact with proteins and other biomolecules, thus forming a dynamic corona whose composition varies over time due to continuous protein association and dissociation events. Eventually equilibrium is reached, at which point the continued exchange will not affect the composition of the corona. Results We developed a simple and effective dynamic model of the nanoparticle protein corona in a body fluid, namely human plasma. The model predicts the time evolution and equilibrium composition of the corona based on affinities, stoichiometries and rate constants. An application to the interaction of human serum albumin, high density lipoprotein (HDL) and fibrinogen with 70 nm N-iso-propylacrylamide/N-tert-butylacrylamide copolymer nanoparticles is presented, including novel experimental data for HDL. Conclusions The simple model presented here can easily be modified to mimic the interaction of the nanoparticle protein corona with a novel biological fluid or compartment once new data will be available, thus opening novel applications in nanotoxicity and nanomedicine.

Structural Changes in Apolipoproteins Bound to Nanoparticles

Nanoparticles are widely used in the pharmaceutical and food industries, but the consequences of exposure to the human body have not been thoroughly investigated. Apolipoprotein A-I (apoAI), the major protein in high-density lipoprotein (HDL), and other lipoproteins are found in the corona around many nanoparticles, but data on protein structural and functional effects are lacking. Here we investigate the structural consequences of the adsorption of apoAI, apolipoprotein B100 (apoB100), and HDL on polystyrene nanoparticles with different surface charges. The results of circular dichroism, fluorescence spectroscopy, and limited proteolysis experiments indicate effects on both secondary and tertiary structures. Plain and negatively charged nanoparticles induce helical structure in apoAI (negative net charge) whereas positively charged nanoparticles reduce the amount of helical structure. Plain and negatively charged particles induce a small blue shift in the tryptophan fluorescence spectrum, which is not noticed with the positively charged particles. Similar results are observed with reconstituted HDL. In apoB100, both secondary and tertiary structures are perturbed by all particles. To investigate the generality of the role of surface charge, parallel experiments were performed using human serum albumin (HSA, negative net charge) and lysozyme (positive net charge). Again, the secondary structure is most affected by nanoparticles carrying an opposite surface charge relative to the protein. Nanoparticles carrying the same net charge as the protein induce only minor structural changes in lysozyme whereas a moderate change is observed for HSA. Thus, surface charge is a critical parameter for predicting structural changes in adsorbed proteins, yet the effect is specific for each protein.

Delivery success rate of engineered nanoparticles in the presence of the protein corona: a systems-level screening

UNLABELLEDnNanoparticles (NPs) for medical applications are often introduced into the body via intravenous injections, leading to the formation of a protein corona on their surface due to the interaction with blood plasma proteins. Depending on its composition and time evolution, the corona will modify the biological behavior of the particle. For successful delivery and targeting, it is therefore important to assess on a quantitative basis how and to what extent the presence of the corona perturbs the specific interaction of a designed NP with its cellular target. We present a theoretical systems-level analysis, in which peptides have been covalently coupled to the surface of nanoparticles, describing the delivery success rate in varying conditions, with regard to protein composition of the surrounding fluid. Dynamic modeling and parameter sensitivity analysis proved to be useful and computationally affordable tools to aid in the design of NPs with increased success rate probability in a biological context.nnnFROM THE CLINICAL EDITORnThe formation of a protein corona consisting of blood plasma proteins on the surface of intravenously delivered nanoparticles may modify the biological behavior of the particles. This team of investigators present a theoretical systems-level analysis of this important and often neglected phenomenon.

Improved oscillator strengths and wavelengths for Os I and Ir I, and new results on early r-process nucleosynthesis ?

The radioactive decay of 238 Ua nd 232 Th has recently been used to determine ages for some of the oldest stars in the Universe. This has highlighted the need for accurate observational constraints on production models for the heaviest r-process elements which might serve as stable references, notably osmium and iridium. In order to provide a firmer basis for the observed abundances, we have performed laser-induced fluorescence measurements and Fourier Transform Spectroscopy to determine new radiative lifetimes and branching fractions for selected levels in Os I and Ir I. From these data, we determine new absolute oscillator strengths and improved wavelengths for 18 Os I and 4 Ir I lines. A reanalysis of VLT spectra of CS 31082-001 and new results for other stars with Os and Ir detections show that (i): the lines in the UV and 4260 A yield reliable Os abundances, while those at 4135, 4420 A are heavily aected by blending; (ii): the Os and Ir abundances are identical in all the stars; (iii): the heavy-element abundances in very metal-poor stars conform closely to the scaled solar r-process pattern throughout the range 56 Z 77; and (iv): neither Os or Ir nor any lighter species are suitable as reference elements for the radioactive decay of Th and U.

Mathematical modeling of the protein corona: implications for nanoparticulate delivery systems

This article discusses the role of the protein corona in delivery systems with tagged nanoparticles and how knowledge of the protein corona can help in optimizing delivery. The basic question is whether and how the binding of proteins and other biomolecules at the nanoparticle surface interfere with the interaction between a tag and its receptor. This is an interesting problem in many respects, but most intriguing are the observed differences in delivery efficiency in vivo compared with protein-free in vitro conditions. In order to understand possible situations that the nanoparticle will face in a protein-rich biological environment, we will first describe the formation of a protein corona and thereafter discuss potential perturbations of the delivery systems when moving from in vitro testing to in vivo applications. We emphasize the role of mathematical modeling in optimizing the design of functionalized nanoparticles to achieve high success of delivery.

The nanoparticle protein corona formed in human blood or human blood fractions

The protein corona formed around nanoparticles in protein-rich fluids plays an important role for nanoparticle biocompatibility, as found in several studies during the last decade. Biological fluids have complex compositions and the molecular components interact and function together in intricate networks. Therefore, the process to isolate blood or the preparation of blood derivatives may lead to differences in the composition of the identified protein corona around nanoparticles. Here, we show distinct differences in the protein corona formed in whole blood, whole blood with EDTA, plasma, or serum. Furthermore, the ratio between particle surface area to protein concentration influences the detected corona. We also show that the nanoparticle size per se influences the formed protein corona due to curvature effects. These results emphasize the need of investigating the formation and biological importance of the protein corona in the same environment as the nanoparticles are intended for or released into.

Improved oscillator strengths and wavelengths in Hf II, with applications to stellar elemental abundances

Aims. We present new and improved radiative lifetimes for eight levels in HfI and 18 levels in HfII, along with oscillator strengths and wavelengths for 195 transitions in HfII. With these data we determine the abundance of hafnium in two chemically peculiar stars: the hot-Am star HR 3383 and the HgMn star chi Lupi, and discuss the implications of the new data to the hafnium abundance for the Sun and the metal-poor galactic halo stars CS 22892-052 and CS 31082-001. Methods. The oscillator strengths are derived by combining radiative lifetimes measured with the laser induced fluorescence technique and branching fractions determined from intensity calibrated Fourier transform spectra. The hafnium abundance in the two sharp-lined peculair stars is determined by comparison of spectra obtained from instruments onboard the Hubble Space Telescope with synthetic spectra, while the abundance of hafnium in the solar photosphere and the metal-poor halo stars is discussed in terms of rescaling previous investigations using the new g f values. Results. The abundance enhancement of hafnium has been determined in HR 3383 to be +1.7 dex and that for chi Lupi A is +1.3 dex. In the course of the analysis we have also determined an abundance enhancement for molybdenum in HR 3383 to be +1.2 dex, which is similar to that known for. Lupi A. The abundances in the metal-poor halo stars CS 31082-001 and CS 22892-052 were rescaled to log epsilon(Hf) = -0.75 and -0.82 respectively, with smaller 1 sigma uncertainty. This has the effect of improving the theoretical fits of r-process nucleosynthesis to abundance data for heavy elements. The change of g f values also implies that the hafnium abundance in the solar photosphere should be reduced by up to 0.2 dex, thereby inducing a discrepancy with the meteoritic hafnium abundance. (Less)

Size and surface chemistry of nanoparticles lead to a variant behavior in the unfolding dynamics of human carbonic anhydrase

The adsorption induced conformational changes of human carbonic anhydrase I (HCAi) and pseudo wild type human carbonic anhydrase II truncated at the 17th residue at the N-terminus (trHCAii) were studied in presence of nanoparticles of different sizes and polarities. Isothermal titration calorimetry (ITC) studies showed that the binding to apolar surfaces is affected by the nanoparticle size in combination with the inherent protein stability. 8-Anilino-1-naphthalenesulfonic acid (ANS) fluorescence revealed that HCAs adsorb to both hydrophilic and hydrophobic surfaces, however the dynamics of the unfolding at the nanoparticle surfaces drastically vary with the polarity. The size of the nanoparticles has opposite effects depending on the polarity of the nanoparticle surface. The apolar nanoparticles induce seconds timescale structural rearrangements whereas polar nanoparticles induce hours timescale structural rearrangements on the same charged HCA variant. Here, a simple model is proposed where the difference in the timescales of adsorption is correlated with the energy barriers for initial docking and structural rearrangements which are firmly regulated by the surface polarity. Near-UV circular dichorism (CD) further supports that both protein variants undergo structural rearrangements at the nanoparticle surfaces regardless of being hard or soft. However, the conformational changes induced by the apolar surfaces differ for each HCA isoform and diverge from the previously reported effect of silica nanoparticles.